中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
12期
2463-2466
,共4页
孙学溥%孙亮%乔海泉%姜洪池%孙学英
孫學溥%孫亮%喬海泉%薑洪池%孫學英
손학부%손량%교해천%강홍지%손학영
肝移植%免疫耐受%基因治疗%腺相关病毒%血红素加氧酶-1
肝移植%免疫耐受%基因治療%腺相關病毒%血紅素加氧酶-1
간이식%면역내수%기인치료%선상관병독%혈홍소가양매-1
Liver transplantation%Immunological tolerance%Gene therapy%Adeno-associated virus%Heme oxygenase-1
目的 探讨腺相关病毒介导血红素加氧酶-1(AAV-HO-1)诱导大鼠肝移植免疫耐受的有效性及其分子机制.方法 按Kamada二套管法建立大鼠原位肝移植模型.在供肝冷保存阶段分别经门脉灌注磷酸盐缓冲液(PBS)、Empty AAV或AAV-HO-1,孵育2h后行DA→Lewis大鼠原位肝移植,检测移植术后大鼠的中位存活时间(MST);血清中白细胞介素(IL)-2、肿瘤坏死因子(TNF)-α的表达;移植肝中CD4+、CD8+,调节性T细胞(Treg细胞、CD4+ CD25+ Foxp3+)的表达;耐受组脾脏中Treg细胞百分率并对耐受组脾脏行混合淋巴细胞培养(MLC)观察免疫耐受.结果 AAV-HO-1组MST为30 d,显著长于PBS组(11 d)或Empty AAV(12 d)(P<0.01),其中20%存活超过90 d;血清中IL-2、TNF-α的表达明显降低,CD4+和CD8+T细胞的浸润减少,Treg细胞在移植肝中的浸润增加.耐受组脾脏中Treg细胞为6.7%,显著提高,行MLC后几乎无淋巴细胞反应.结论 AAV-HO-1可通过增加Treg细胞,延长大鼠的存活期,证实AAV-HO-1可诱导肝移植免疫耐受.
目的 探討腺相關病毒介導血紅素加氧酶-1(AAV-HO-1)誘導大鼠肝移植免疫耐受的有效性及其分子機製.方法 按Kamada二套管法建立大鼠原位肝移植模型.在供肝冷保存階段分彆經門脈灌註燐痠鹽緩遲液(PBS)、Empty AAV或AAV-HO-1,孵育2h後行DA→Lewis大鼠原位肝移植,檢測移植術後大鼠的中位存活時間(MST);血清中白細胞介素(IL)-2、腫瘤壞死因子(TNF)-α的錶達;移植肝中CD4+、CD8+,調節性T細胞(Treg細胞、CD4+ CD25+ Foxp3+)的錶達;耐受組脾髒中Treg細胞百分率併對耐受組脾髒行混閤淋巴細胞培養(MLC)觀察免疫耐受.結果 AAV-HO-1組MST為30 d,顯著長于PBS組(11 d)或Empty AAV(12 d)(P<0.01),其中20%存活超過90 d;血清中IL-2、TNF-α的錶達明顯降低,CD4+和CD8+T細胞的浸潤減少,Treg細胞在移植肝中的浸潤增加.耐受組脾髒中Treg細胞為6.7%,顯著提高,行MLC後幾乎無淋巴細胞反應.結論 AAV-HO-1可通過增加Treg細胞,延長大鼠的存活期,證實AAV-HO-1可誘導肝移植免疫耐受.
목적 탐토선상관병독개도혈홍소가양매-1(AAV-HO-1)유도대서간이식면역내수적유효성급기분자궤제.방법 안Kamada이투관법건립대서원위간이식모형.재공간랭보존계단분별경문맥관주린산염완충액(PBS)、Empty AAV혹AAV-HO-1,부육2h후행DA→Lewis대서원위간이식,검측이식술후대서적중위존활시간(MST);혈청중백세포개소(IL)-2、종류배사인자(TNF)-α적표체;이식간중CD4+、CD8+,조절성T세포(Treg세포、CD4+ CD25+ Foxp3+)적표체;내수조비장중Treg세포백분솔병대내수조비장행혼합림파세포배양(MLC)관찰면역내수.결과 AAV-HO-1조MST위30 d,현저장우PBS조(11 d)혹Empty AAV(12 d)(P<0.01),기중20%존활초과90 d;혈청중IL-2、TNF-α적표체명현강저,CD4+화CD8+T세포적침윤감소,Treg세포재이식간중적침윤증가.내수조비장중Treg세포위6.7%,현저제고,행MLC후궤호무림파세포반응.결론 AAV-HO-1가통과증가Treg세포,연장대서적존활기,증실AAV-HO-1가유도간이식면역내수.
Objective To investigate the efficacy and molecular mechanisms of AAV-HO-1 inducing immuno-telorance in rats.Methods An orthotopic liver transplantation model by DA to Lewis was set up using Kamada' s two cuff technique.PBS,empty AAV or purified AAV-HO-1 was injected into the portal vein and incubated for 2 h at the donor liver cold preserved stage,and then OLT was done.The media survival time (MST),serum levels of interleukin (IL)-2 and tumor necrosis factor (TNF)-α,infiltration of CD4 +,CD8 + and Treg (CD4 + CD25 + Foxp3 +) cells into donor livers and percent of Treg in the spleen were examined.A mixed lymphocyte reaction (MLR) was performed.Results Recipients transplanted with AAV-HO-1-perfused liver allografts had a median survival time of 30 days,which was significantly longer than that in the PBS group (11 days) and empty AAV group (12 days) (both P <0.01).Overexpression of HO-1 reduced the production of IL-2 and TNF-α,inhibited infiltration of CD4+ and CD8 + cells,and increased infiltration of Treg ceils into donor livers.Splenocytes from the tolerant recipients had higher percentages of Treg cells,and responded poorly to the allogeneic donor splenocytes.Conclusion Persistent expression of HO-1 in the donor livers by intraportal delivery of AAV-HO-1 can prolong the survival by expanding Treg cells,suggesting that AAV-HO-1 can induce immuno-tolerance in liver allograft in rats.